低氧对人骨髓间充质干细胞向神经细胞分化的影响

发布时间：2018-06-21 00:39

本文选题：低氧 + 人骨髓间充质干细胞　；参考：《广东医学》2017年20期

【摘要】：目的探讨低氧对人骨髓间充质干细胞(hB MSCs)向神经细胞分化的影响。方法将第2代hB M-SCs进行传代、纯化,取第4代hB MSCs进行流式细胞仪检测鉴定;以含1%FBS的α-MEM为基础诱导培养基,根据处理条件不同分为4组:常氧对照组、低氧对照组(3%氧浓度)、常氧诱导组[添加20 ng/mL表皮生长因子(EGF)及20 ng/mL碱性成纤维细胞生长因子(bF GF)]、低氧诱导组(3%氧浓度下添加20 ng/mL EGF及20 ng/mL bF GF)。连续诱导3 d后,采用RT-PCR及Western blot技术检测神经元特异性烯醇化酶(NSE)、微管相关蛋白2(MAP-2)及胶质纤维酸性蛋白(GFAP)在基因及蛋白水平表达变化。结果第4代hB MSCs流式鉴定结果示:骨髓基质标志CD105、CD90、CD29表达分别为95.8%、98.2%、95.1%,而造血细胞标志CD19表达为0.2%;与常氧对照组比较,低氧对照组NSE、MAP-2、GFAP在基因及蛋白水平表达均无明显改变(P0.05),而常氧诱导组及低氧诱导组相应表达均升高(P0.05),且低氧诱导组表达高于常氧诱导组(P0.05)。结论低氧促进EGF联合bF GF对hB MSCs向神经元细胞及神经胶质细胞分化的诱导作用。
[Abstract]:Objective to investigate the effect of hypoxia on the differentiation of human bone marrow mesenchymal stem cells (hBMSCs) into neural cells. Methods the second passage of hB M-SCs was subcultured, purified, the fourth passage of hB MSCs was detected and identified by flow cytometry, and on the basis of 伪 -MEM containing 1s, the MSCs were divided into four groups according to different treatment conditions: normoxic control group, normal control group, and normal control group. Hypoxia control group (3% oxygen concentration), normoxia-induced group (20 ng / mL epidermal growth factor EGF) and 20 ng / mL basic fibroblast growth factor (BGF) were added 20 ng / mL EGF and 20 ng / mL BF at 3% oxygen concentration. After induction for 3 days, the expression of neuron-specific enolase (NSEP), microtubule-associated protein (MAP-2) and glial fibrillary acidic protein (GFAP) were detected by RT-PCR and Western blot. Results the expression of bone marrow stromal marker CD105, CD90, CD90, CD29, and hematopoietic cell marker CD19 was 95. 8%, 98. 2% and 0. 2% respectively, compared with that of normoxic control group. In hypoxic control group, the expression of MAP-2GFAP in gene and protein level was not significantly changed, but the expression of P0.05in normoxic and hypoxic groups was increased, and the expression in hypoxia induced group was higher than that in normoxia-induced group. Conclusion hypoxia can induce differentiation of hB MSCs into neuronal cells and glial cells.
【作者单位】：景德镇市第一人民医院骨二科;九江学院转化医学重点实验室;
【基金】：江西省教育厅科学技术研究项目(编号:GJJ13740)
【分类号】：R651.2

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